International Journal of Fauna and Biological Studies 2017; 4(6): 48-54

ISSN 2347-2677 IJFBS 2017; 4(6): 48-54 Received: 29-09-2017 Histopathological studies of selected organs of Accepted: 30-10-2017 Sarotherodon galilaeus (Linnaeus, 1758) in Igun gold

Komolafe Olaniyi Olusola mining reservoir and Opa freshwater reservoir, south Department of Zoology, Obafemi Awolowo University, Ile-Ife, western, Nigeria Nigeria

Obayemi Oluwadamilare Emmanuel Komolafe Olaniyi Olusola, Obayemi Oluwadamilare Emmanuel, Department of Zoology, Obafemi Awodiran Michael Olufemi and Oladimeji Tofunmi Elizabeth Awolowo University, Ile-Ife, Nigeria Abstract Awodiran Michael Olufemi Histological changes in the gills, fillet and liver of Sarotherodon galileaus in Opa and Igun reservoirs Department of Zoology, Obafemi were used to evaluate the impact of heavy metals in the fish organs. Fresh fish samples of S. galileaus Awolowo University, Ile-Ife, were collected from Opa and Igun reservoirs and identified in the laboratory. Techniques based on Nigeria histological analyses were done on the tissues and photomicrographs taken using digital binocular compound LED microscope. In Opa and Igun reservoirs, the gills of S. galilaeus showed rupture and Oladimeji Tofunmi Elizabeth lifting of epithelium. The gills also showed shortening and hyperplasia of lamellae in Igun reservoir Department of Zoology, Obafemi compared to Opa reservoir. The fillet of S. galilaeus in both reservoirs revealed muscular atrophy and Awolowo University, Ile-Ife, splitting while necrosis was observed in Igun reservoir compared to Opa reservoir. The liver of S. Nigeria galilaeus in the two reservoirs showed hepatopancreas degeneration while splitting at the wall of central

vein, hypertrophy, degeneration and vascular congestion were observed only in the liver of S. galilaeus

of Igun reservoir. The alterations were comparatively severe in the organs of S. galilaeus in Igun reservoir to Opa reservoir.

Keywords: Sarotherodon galilaeus, Histopathology, Reservoirs, Opa, Igun

Introduction The mango also known as Sarotherodon galilaeus (Linnaeus, 1758) is a fish species

that belong to the family. This fish species is found in lakes, rivers and other fresh water bodies in northern, central and western Africa including Nigeria. Histopathology is the microscopic assessment of altered or diseased structure as described by [1]. [2] noted that the study of histopathology of fish is of main importance in the diagnosis, etiology and prevention of disease. Therefore, fish are commonly considered as organism for assessing the impacts of [3] environmental contamination on aquatic biota . Also, Fish are very vulnerable to [4] environmental changes and respond significantly to aquatic pollution . Histological study provides a rapid process to detect effects of pollutants in various organs and tissues of fish [5]. Similarly, it is useful to have an insight into histological investigation, as they serve as biological indicators to evaluate the toxicity condition of fish [6] [7]. [8] noted that fish are usually use to evaluate the health of aquatic biota because contaminants build up in the food chain may be responsible for adverse effects and death in the aquatic life. Also, water borne metals may alter the biochemical and physiological parameters in fish tissues as described by [9]. The authors were of the view that these heavy metals cannot be destroyed through biological and may cause histological alterations in tissues. The most common sources of heavy metal contamination are mining activities, industrial exploration [10] processing and sewage disposal . Pollution of water bodies with a wide range of contaminants has become a matter of concern [11]. This is because heavy metals discharged from domestic, industrial and other man made activities may pollute the water bodies extensively [12]. Also, heavy metals accumulated in the fish tissues may lead to morphological alterations in the fish [13]. The liver is associated with Correspondence Komolafe Olaniyi Olusola the detoxification and biotransformation process. It is also one of the tissues affected by [14] Department of Zoology, Obafemi pollutants in the aquatic environment . Similarly, the gills are responsible for respiration and Awolowo University, Ile-Ife, transport system involved in osmoregulation. It has also been established that bioaccumulation Nigeria of metal ions within them may affect their functions [15]. The gills and skin are primary ~ 48 ~ International Journal of Fauna and Biological Studies

Markers for aquatic pollution because they exhibited large State and Opa freshwater reservoir at Ife central local surfaces which are in direct and permanent contact with Government area of Osun State. The abandoned gold mine potential irritants. Several Studies have been conducted on reservoir extends over longitude 004030E-004045E and histopathological changes in the gills, liver and fillet of fish latitude 07035N-07038N (figure 1). Streams such as Oika, species exposed to various chemicals including pesticide Eleripon and Osun which serve the community were which have been reported to cause pathological alterations [16]. impounded to form reservoirs in order to meet the mining Therefore, the aim of the present study is to evaluate the needs of the Nigerian Mining cooperation which started in occurrence of histopathological alterations in the gills, fillet December 1941. and liver of Sarotherodon galileaus of Opa and Igun The second study area which is Opa reservoir was impounded reservoirs. This is as a result of reported high levels of heavy in 1978. The major tributaries are rivers Opa, Obudu and metals in the fish species of Igun reservoir [17] when compared Esinmirin. The reservoir has a catchment area of about with that of Opa reservoir [18]. 116km. River Opa is a stream and is located in Ile-Ife, Osun state Nigeria (figure 1). The estimated terrain elevation Materials and methods Above sea level is 196 meter. The reservoir extends over Study area latitudes 07021'N and 07035'N and longitudes 004031'E and The study areas are abandoned gold mine reservoir at Igun 004039'E. village in Atakunmosa West Local Government area of Osun

Fig 1: Map of Opa and Igun reservoirs showing its location in Nigeria

Collection of fish samples cut into sections of approximately 5 µm thickness from the Fish samples were collected on a monthly basis using cast net block using a rotary microtome (Yamato Kohki, Serial no: between August 2015 and July 2016. Fishes were identified 75010JO). The cut samples were dried in a hot air oven to using standard keys prepared by [19] and [20]. Samples of fish remove moisture and each section were mounted on a glass caught were put in a container filled with the reservoir water slide. The sections were de-waxed in a wax-miscible agent, and dissected in situ. rehydrated through descending concentrations of ethanol (90%, 80% and 70% alcohol) for at least 2 minutes. The Preparation of fish tissues and organs for Histological sections were then stained with haematoxylin and eosin [21], in analysis which the tissues were place in haematoxylin solution for 3 Each fish specimen was split open anteriorly from the anal minutes and aqueous eosin for 3 minutes, mounted on a slide pore to the pectoral fin to remove its liver, while the gills and covered with coverslip and labelled appropriately. The were removed from the head region. A piece of fillet was also tissues were examined, and microphotographs taken using a taken just above the lateral line and before the dorsal fin. Each digital binocular compound LED microscope (model fish gills, fillet and liver were put in a separate well labelled MD827S30L series). bottle, fixed in 5% formalin for at least 48 hours and transferred into a sampling bottle rack. The method of [5] was Results used for tissues processing for histological studies, the tissues Histopathological alterations in the organs of were removed from the fixative, and samples of tissue were Sarotherodon galilaeus in Opa and Igun reservoirs rinsed in tap water for 5 minutes, dehydrated in ascending Histopathological alterations observed in the gills of S. ethanol concentrations (70%, 80% and 90% alcohol) for galilaeus of Opa reservoir are curling of secondary lamellae minimum of 2 minutes, cleared or infiltrated in a wax (Figure 2.1a), lifting at the tip of secondary lamellae, miscible agent (xylene) for 2 minutes and then embedded in epithelial lifting, rupture of gill epithelium (Figure 2.1e). paraffin using standard protocols. The fish tissues were then Also, abnormalities in the gills of S. galilaeus in Igun

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reservoir include epithelial lifting, shortening of secondary lamellae (Figure 2.1d). Similarly, Figure 2.1f showed hyperplasia of secondary lamellae, rupture of chloride cell and rupture of gill epithelium. Lifting and rupture of epithelium were observed in the gills of S. galilaeus in the two reservoirs. Also, curling and lifting of lamellae were identified in the fish of Opa reservoir when compared to rupture of chloride cell, shortening and hyperplasia of lamellae observed in the gills of S. galialeus of Igun reservoir. Similarly, the alterations revealed in the fillet of S. galilaeus in Opa reservoir are atrophy of muscle bundles (Figure 2.2c), splitting of muscle myofibril (Figure 2.2e). Further, the fillet alterations in S. galilaeus of Igun reservoir include atrophy of muscle bundles (Figure 2.2b), necrosis of muscle bundles in Fig 2.1c: Photomicrograph of gill section Figure 2.2d, splitting of muscle myofibrils and muscle bundles (Figure 2.2f). Atrophy and splitting of muscle myofibrils were observed in the gills of the fish in Opa amd Igun reservoirs. In contrast, the fillet of S. galilaeus exhibited histopathological changes such as necrosis and splitting of muscle bundles. Also, the pathological alterations observed in the liver of S. galilaeus in Opa reservoir are vascular congestion in central vein (Figure 2.3a), hepatopancreas degeneration (Figure 2.3c). Histopathological alterations observed in the liver of S. galilaeus from Igun reservoir include spltting at the wall of central vein, degeneration of liver cells (Figure 2.3b), vascular congestion in portal vein and hepatopancreas degeneration in Figure 2.3d. Nucleus hypertrophy and vascular congestion in sinusoids was also observed as shown in (Figure 2.3f).

Fig 2.1d: Photomicrograph of gill section in S. galilaeus of Opa reservoir (Mag. X400) in S. galilaeus of Igun reservoir (Mag. X400)

Fig 2.1a: Photomicrograph of gill section

Fig 2.1e: Photomicrograph of gill section

Fig 2.1b: Photomicrograph of gill section in Sarotherodon galilaeus

of Opa reservoir in S. galilaeus of Igun reservoir (Mag. X100) (Mag. Fig 2.1f: Photomicrograph of gill section in in S. galilaeus of Opa X100) reservoir (Mag. X400) in S. galilaeus of Igun reservoir (Mag. X400)

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Rainy season keys: Curling of secondary lamellae (CSL), primary lamellae (PL), secondary lamellae (SL), epithelial cell (EC), chloride cell (CC), shortening of the secondary lamellae (SSL), epithelial lifting (EL), lifting at the tip of the secondary lamellae (LSL), mucous cell (MC), rupture of chloride cell (RCC) hyperplasia of secondary lamellae (HSL) and rupture of gill epithelium (RGE). Haematoxylin and Eosin stain.

Fig 2.2d: Photomicrograph of fillet section in S. galilaeus of Opa reservoir (Mag. X40) in S. galilaeus of Igun reservoir (Mag. X40)

Fig 2.2a: Photomicrograph of fillet section

Fig 2.2e: Photomicrograph of fillet section

Fig 2.2f: Photomicrograph of fillet section in S. galilaeus of Opa Fig 2.2b: Photomicrograph of fillet section in Sarotherodon reservoir (Mag. X100) in S. galilaeus of Igun reservoir (Mag. X100) galilaeus of Opa reservoir in S. galilaeus of Igun reservoir (Mag. X40) (Mag. X40) Rainy season keys: normal arrangement of muscle bundles (NAM), atrophy of muscle bundles (AMB), necrosis of muscle bundles (NMB), splitting of muscle myofibrils (SMM) and splitting of muscle bundles (SMB). Haematoxylin and Eosin stain.

Fig 2.2c: Photomicrograph of fillet section

Fig 2.3a: Photomicrograph of liver section ~ 51 ~ International Journal of Fauna and Biological Studies

Fig 2.3b: Photomicrograph of liver section in Sarotherodon galilaeus of Opa reservoir in S. galilaeus of Igun reservoir (Mag. Fig 2.3f: Photomicrograph of liver section in S. galilaeus of Opa X40) (Mag. X40) reservoir (Mag. X400) in S. galilaeus of Igun reservoir (Mag. X400)

Rainy season keys: central vein (CV), vascular congestion (VC), splitting at the wall of central vein (SCV), degeneration of liver cells (DLC), nucleus (N), hepatocytes (H), nucleus hypertrophy (NH), vascular congestion in sinusoids (VCS), portal vein (PV), hepatopancreas (HP), bile duct (BD), potal artery (PA) and hepatopanreas degeneration (HD). Haematoxylin and Eosin stain.

Discussion The rupture of gill epithelium observed in the gills of S. galiaeus in Opa and Igun reservoirs was similar to the review by [22]. According to the review on alterations induced by

toxic substances in fish gills, the most common Fig 2.3c: Photomicrograph of liver section histopathological changes were hyperplasia, hypertrophy and rupture of gill epithelium. Also, epithelial lifting, curling and lifting of secondary lamellae observed in the gills of S. galilaeus in Opa reservoir were similar to the results obtained by [23] that reported histopathological changes in gills and liver of Chana punctatus. The lifting of lamellar epithelium could probably be induced by incidence of severe edema according to [24]. Hence, it could be suggested that there is an increasing level of pollution in the two reservoirs. Similarly, shortening of secondary lamellae, hyperplasia of secondary lamellae and rupture of chloride cells were revealed in the gills of S. galilaues in Igun reservoir are also in agreement with the result of [23] on Channa punctatus under laboratory condition. Hyperplasia of lamellae which is a

Fig 2.3d: Photomicrograph of liver section in S. galilaeus of Opa common responses, is more common for metals than for reservoir (Mag. X100) in S. galilaeus of Igun reservoir (Mag. X100) organics or other pollutants, possibly since metals directly interact with ion transport proteins and inhibit their activity according to [25]. Also, alterations found in the fillet of S. galilaeus in Opa and Igun reservoirs were atrophy of muscle bundles and splitting of muscle bundles. This finding is in accordance with the report of [26] who recorded histopathological alterations in Danio rerio under laboratory condition. Splitting of muscle myofibrils and necrosis of muscle bundles found in the fillet of S. galilaeus in Igun reservoir were similar with alterations recorded by [27] (Begum et al., 2013) on Heteropneustes fossilis expose to Arsenic. This result may be explained by the fact that pollution may affect changes in fish tissues. Furthermore, the liver of S. galilaeus in both Opa and Igun reservoirs showed hepatopancreas degeneration. This condition refers to reduction in cell size. Vascular congestion in central vein was observed in the liver of S. galilaeus of Opa Fig 2.3e: Photomicrograph of liver section reservoir. Alterations showed in the liver of S. galilaeus in ~ 52 ~ International Journal of Fauna and Biological Studies

Igun reservoir corroborates the work of [28]. These are and Technology. 2008; 5(2):174-182. degeneration of liver cells, splitting at the wall of central vein, 10. Dimari GA, Abdulkarim FI, Akan, JC, Garba ST. vascular congestion in sinusoids and portal vein. Cellular Metal concentrations in Tissues of Tilapia galier, degeneration and necrosis may be due to accumulative effect Clarias lazera, and Osteoglosidae caught from Alau of metals in hepatic tissue according to [27]. Dam, Maiduguri, Borno State, Nigeria. American Journal of Environmental Sciences. 2008; 4(4):473- Conclusion 379. The investigation of the organs of S. galilaeus in Igun 11. Vutukuru SS. Acute effect of hexavalent chromium on reservoir compared to Opa reservoir has shown that more survival, oxygen consumption, haematological alterations were found in the organs of S. galileaus in Igun parameters and some biochemical profiles of the reservoir. 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